A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax

Kagawa, Shunsuke; Pearson, Sally-Ann; Ji, Lin; Xu, Kang; McDonnell, Timothy J.; Swisher, Stephen G.; Roth, Jack A.; Fang, Bingliang

doi:10.1038/sj.gt.3301048

Cited by 82 publications

(78 citation statements)

References 14 publications

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“…Histopathologic examination of liver sections of mice systemically treated with iodixanol-purified Ad/RGDLacZ at doses between 1 × 10 11 to 1 × 10 12 VP showed changes that are commonly observed in animals treated with E-deleted adenovectors, including mild portal triaditis, mild hepatocellular degeneration (cytomegaly and karyomegaly), and apoptosis in 1-3% of hepatocytes. Similar histopathologic changes have been observed in animals treated with CsClpurified adenovectors at doses of less than 1 × 10 11 VP [24,26], suggesting that adenovectors purified by iodixanol are at least as safe as those purified by the CsCl method. …”

supporting

confidence: 58%

A rapid and efficient method for purification of recombinant adenovirus with arginine–glycine–aspartic acid-modified fibers

Peng¹,

Wu²,

Davis³

et al. 2006

Analytical Biochemistry

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Recombinant adenoviral vectors (adenovectors) have been subject to various genetic modifications to improve their transduction efficiency and targeting capacity. Production and purification of adenovectors with modified capsid proteins can be problematic using conventional two-cycle CsCl gradient ultracentrifugation. We have developed a new method for purifying recombinant adenovectors in two steps: iodixanol discontinuous density gradient ultracentrifugation and sizeexclusion column chromatography. The purity and infectious activity of adenovectors isolated by either method were comparable. The new method yielded three to four times more adenovectors with RGD-modified fiber proteins than did the conventional CsCl method. For other fiber-modified and wild-type adenovectors, the yields of the two methods were comparable. Thus, the iodixanol-based method can be used not only to improve the production of RGD-modified adenovectors, but also to purify adenovectors with or without fiber modifications. Moreover, the whole procedure can be completed in 3 hours. Therefore, this method is rapid and efficient for production recombination adenovectors, especially those with RGD-modified fibers.Adenoviral vectors (adenovectors) have high in vivo transduction efficiencies and are easy to construct and produce. They are thus frequently used for in vitro and in vivo gene delivery and for gene therapy in clinical trials [1] . Substantial efforts have been made to characterize adenovirus-host cell interactions and to improve gene delivery by adenovectors, including minimizing vector gene expression, reducing vector related toxicity and immunogenicity, increasing their capacity to accommodate large foreign genes, and increasing their transduction efficiency.It is now known that adenovirus interacts with host cells by binding to the coxsackievirus and adenovirus receptor (CAR) [2], a cellular receptor for Ad5 and most other adenovirus serotypes. After attachment, the virion is then internalized by endocytosis through the interaction of its penton base with α v β 3 and α v β 5 integrins on the host cell surface [3]. However, there is growing evidence that the expression of CAR is frequently suppressed in various types of cells, including tumor cells and in primary tumors [4,5], resulting in resistance to adenovirus infection [6][7][8] Unfortunately, technical difficulties may be encountered in production of certain modified adenovectors. We have encountered difficulties when producing certain adenovectors by this method, especially those with RGD-modified fibers or "sticky" virus. For example, we have experienced difficulty in purifying various RGD-modified adenoviral vectors by conventional two-cycle (2x) of CsCl ultracentrifugation because the RGD-modified adenoviral vectors tend to aggregate, especially in the second round of ultracentrifugation, forming floccules without a sharp band. This results in a low yield or even failure of purification. Thus, an alternative method for efficient production of these modified adenov...

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supporting

confidence: 58%

A rapid and efficient method for purification of recombinant adenovirus with arginine–glycine–aspartic acid-modified fibers

Peng¹,

Wu²,

Davis³

et al. 2006

Analytical Biochemistry

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“…5,22 Seventy-two hours after infection, about 95% of cells in each treatment were apoptotic. In contrast, treatment with a control vector Ad/CMV-GFP at the same total dose resulted in only background levels of cell death that were similar to the level of apoptpsis seen in mock control.…”

Section: Selection Of Dld1/bax-r and Dld1/trail-r Cellsmentioning

confidence: 99%

Mechanisms involved in development of resistance to adenovirus-mediated proapoptotic gene therapy in DLD1 human colon cancer cell line

Zhang

Lin

et al. 2002

Gene Ther

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“…Previously, for example, it has been shown that systemic administration of adenovirus vectors expressing the Bax gene suppressed tumor growth in vivo but also resulted in massive apoptosis in the liver. 27 Although systemic delivery was not attempted in the present studies, the present results suggest that transcriptional targeting through the use of SCCA2 promoter may prove useful in reducing the potential for adverse side effects in normal tissues and in enhancing the therapeutic effect in SCC tumors. To prove the safety certainly, the systemic injection of the vectors into mice and the analyses of major organs will be necessary and subject in future.…”

Section: Discussionmentioning

confidence: 73%

Transcriptional targeting of adenovirus vectors with the squamous cell carcinoma-specific antigen-2 promoter for selective apoptosis induction in lung cancer

et al. 2006

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Squamous cell carcinoma antigens SCCA1 and SCCA2 are highly homologous serine proteinase inhibitors which have been widely utilized as serological markers for squamous cell cancers, but it has recently been demonstrated that only SCCA2 is truly specific for certain forms of lung cancer. Using a construct containing the 5 0 -flanking region of the SCCA2 gene between À460 and þ 0 bp and the luciferase reporter gene, SCCA2 promoter activity was detected in SCCA2-producing SCC cell lines (LK-2, LC-1), but not in SCCA2-nonproducing lung adenocarcinoma cell lines (A549, ABC-1, and RERF-LC-MS) or normal cells (WI-38, SAEC, and NHEKAdult). Infection with a recombinant adenovirus vector, Ad-SCCA2-DsRed, resulted in cell-specific expression of the SCCA2 promoter-driven DsRed marker gene only in LK-2 and LC-1 cells. The same strategy was used for SCCA2-driven expression of a proapoptotic gene, (KLAKLAK)2, which can cause mitochondrial disruption by triggering mitochondrial permeabilization and swelling, resulting in the release of cytochrome c and induction of apoptosis. Infection with Ad-SCCA2-KLAKLAK2 specifically reduced the growth of the two human lung SCC cell lines compared to the SCCA2 nonproducing cell lines both in vitro and in vivo, suggesting that the SCCA2 promoter had a tumor-specific effect. These results suggest that transduction of SCCA2 promotercontrolled suicide genes by adenoviral vectors can confer transcriptionally targeted cytotoxicity in SCCA2-producing lung SCC cells, and represents a novel strategy for gene transfer specifically targeted to SCC in the lung. Cancer Gene Therapy ( IntroductionLung carcinoma is the most common malignant neoplasm and represents the leading cause of cancer death in the developed world. 1 Among lung carcinomas, adenocarcinoma and squamous cell carcinoma (SCC) are the most frequently encountered forms. Despite the use of optimal chemotherapy and radiotherapy regimens, there has been little improvement in the prognosis of these cancer patients during the last decade. 2,3 This underscores the need to seek novel therapeutic modalities for lung cancer patients. One such approach is gene therapy by administration of vectors expressing cytotoxic genes; however, in order to maximize tumor-killing efficacy and minimize any potential adverse effects of this approach, it would be advantageous to employ a cancer-specific targeting strategy. For adenocarcinomas in general, transcriptional targeting by the use of cancer-specific promoters such as the carcinoembryonic antigen (CEA) promoter has been described previously. 4,5 However, little has been reported for SCC-specific regulation.The squamous cell carcinoma antigen (SCCA) was first reported by Kato et al. 6 in 1977 as a circulating tumor marker for SCC, particularly that of cervix, head and neck, esophagus, and lung. 7 Several studies have shown that increased serum SCCA levels are seen in SCC patients 8,9 and correlate with the extent of disease in patients with SCC, 10,11 although the protein itself is not detected in adenoca...

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A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax

Cited by 82 publications

References 14 publications

A rapid and efficient method for purification of recombinant adenovirus with arginine–glycine–aspartic acid-modified fibers

A rapid and efficient method for purification of recombinant adenovirus with arginine–glycine–aspartic acid-modified fibers

Mechanisms involved in development of resistance to adenovirus-mediated proapoptotic gene therapy in DLD1 human colon cancer cell line

Transcriptional targeting of adenovirus vectors with the squamous cell carcinoma-specific antigen-2 promoter for selective apoptosis induction in lung cancer

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